Cardiac Myocyte Hypertrophy Research Articles

Abstract P2005: Antioxidant Resveratrol Attenuates Cardiac Fibrosis and Reduces Matrix Metalloproteinase-2 Activity in Heart Failure Rats

Increased reactive oxygen species activate matrix metalloproteinases (MMP) playing a critical role in cardiovascular remodeling after myocardial infarction (MI). We evaluated the antioxidant effect of resveratrol (RESV) on cardiac remodeling and MMP-2 activity in heart failure (HF) rats. Wistar rats were divided into the following groups: Control; Control+RESV; HF; HF+RESV (10mg/kg/day, p.o., 4 wk). The MI was elicited by ligation of the coronary artery. After four weeks, the animals were anesthetized to evaluate the cardiac function and to collect the hearts and blood samples. Heart weight/body weight (HW/BW, mg/g) ratio and the infarct size were calculated. Histological assessment of the left ventricle was carried out with hematoxylin/eosin, and fibrosis was quantified with Masson trichrome-staining. Cardiac and plasma MMP-2 activities were determined by gel zymography. The oxidative stress was assessed measuring the thiobarbituric acid reactive substances (TBARS) from plasma. HF and HF+RESV showed similar infarct size. The HW/BW ratio increased (3.7±0.2 vs 3.8±0.2 vs 3.0±0.05). RESV improved the maximal slope of the systolic increment of left ventricular pressure in HF animals (9004±1120 vs 5849±595 vs 7181±533 mmHg/s) and reduced the end diastolic pressure (4±1 vs 18±4 vs 7±1 mmHg). HF increased TBARS, whereas RESV produced an antioxidant effect in HF+RESV group (11.2±0.9 vs 21.7±3 vs 8.2±1.4 μmol/L). HF augmented cardiac fibrosis (30,8±1,3 vs 8,8±0,5%) and myocyte hypertrophy (12,2±0,7 vs 8,8±0,3μm). RESV reduced collagen deposition in HF hearts (22,7±1,9%), but did not reduce the myocyte hypertrophy (11,7±0,6 μm). Cardiac and plasma MMP-2 activity were higher in HF animals (plasma: 1344±39 vs. 1164±47; heart: 536±54 vs. 262±18). RESV decreased plasma MMP-2 activity in control (992±27) and HF animals (1200±49) and reduced cardiac MMP-2 activity only in HF+RESV group (426±42). In conclusion, the antioxidant effect of RESV improved cardiac function and fibrosis and reduced MMP-2 levels in the heart and plasma from HF rats.

Golgi localized β1-adrenergic receptors stimulate Golgi PI4P hydrolysis by PLCε to regulate cardiac hypertrophy.

Increased adrenergic tone resulting from cardiovascular stress leads to development of heart failure, in part, through chronic stimulation of β1 adrenergic receptors (βARs) on cardiac myocytes. Blocking these receptors is part of the basis for β-blocker therapy for heart failure. Recent data demonstrate that G protein-coupled receptors (GPCRs), including βARs, are activated intracellularly, although the biological significance is unclear. Here we investigated the functional role of Golgi βARs in rat cardiac myocytes and found they activate Golgi localized, prohypertrophic, phosphoinositide hydrolysis, that is not accessed by cell surface βAR stimulation. This pathway is accessed by the physiological neurotransmitter norepinephrine (NE) via an Oct3 organic cation transporter. Blockade of Oct3 or specific blockade of Golgi resident β1ARs prevents NE dependent cardiac myocyte hypertrophy. This clearly defines a pathway activated by internal GPCRs in a biologically relevant cell type and has implications for development of more efficacious β-blocker therapies.

Diagnóstico y estudio de cardiopatías infrecuentes: multimodalidad – miocardiopatía hipertrófica

Hypertrophic cardiomyopathy is a genetic disorder, characterised by left ventricular hypertrophy not explained by secondary causes, including non-dilated ventricle and preserved or augmented ejection fraction.1 Histologically, hypertrophic cardiac myocytes are observed that are disorganised, losing parallel alignment due to random orientation, and separated by wide areas of fibrosis.The estimated prevalence in the adult population is 1:625 – 1:344, no ethic, geographic or sex distribution patterns have been observed.2 Ventricular hypertrophy is the cornerstone in the diagnosis of hypertrophic cardiomyopathy. The presence of a ventricular wall greater than 15mm, suffices initially to consider the possibility of hypertrophic cardiomyopathy.The different imaging modalities in cardiology such as echocardiogram and cardiac resonance are fundamental in the diagnosis of the entity, and when determining prognosis and assessing response to treatment.

A high-throughput ratiometric method for imaging hypertrophic growth in cultured primary cardiac myocytes

Maladaptive hypertrophy of cardiac myocytes increases the risk of heart failure. The underlying signaling can be triggered and interrogated in cultured neonatal ventricular myocytes (NRVMs) using sophisticated pharmacological and genetic techniques. However, the methods for quantifying cell growth are, by comparison, inadequate. The lack of quantitative, calibratable and computationally-inexpensive high-throughput technology has limited the scope for using cultured myocytes in large-scale analyses. We present a ratiometric method for quantifying the hypertrophic growth of cultured myocytes, compatible with high-throughput imaging platforms. Protein biomass was assayed from sulforhodamine B (SRB) fluorescence, and image analysis calculated the quotient of signal from extra-nuclear and nuclear regions. The former readout relates to hypertrophic growth, whereas the latter is a reference for correcting protein-independent (e.g. equipment-related) variables. This ratiometric measure, when normalized to the number of cells, provides a robust quantification of cellular hypertrophy. The method was tested by comparing the efficacy of various chemical agonists to evoke hypertrophy, and verified using independent assays (myocyte area, transcripts of markers). The method's high resolving power and wide dynamic range were confirmed by the ability to generate concentration-response curves, track the time-course of hypertrophic responses with fine temporal resolution, describe drug/agonist interactions, and screen for novel anti-hypertrophic agents. The method can be implemented as an end-point in protocols investigating hypertrophy, and is compatible with automated plate-reader platforms for generating high-throughput data, thereby reducing investigator-bias. Finally, the computationally-minimal workflow required for obtaining measurements makes the method simple to implement in most laboratories.

ATF6 Regulates Cardiac Hypertrophy by Transcriptional Induction of the mTORC1 Activator, Rheb.

Endoplasmic reticulum (ER) stress dysregulates ER proteostasis, which activates the transcription factor, ATF6 (activating transcription factor 6α), an inducer of genes that enhance protein folding and restore ER proteostasis. Because of increased protein synthesis, it is possible that protein folding and ER proteostasis are challenged during cardiac myocyte growth. However, it is not known whether ATF6 is activated, and if so, what its function is during hypertrophic growth of cardiac myocytes. To examine the activity and function of ATF6 during cardiac hypertrophy. We found that ER stress and ATF6 were activated and ATF6 target genes were induced in mice subjected to an acute model of transverse aortic constriction, or to free-wheel exercise, both of which promote adaptive cardiac myocyte hypertrophy with preserved cardiac function. Cardiac myocyte-specific deletion of Atf6 (ATF6 cKO [conditional knockout]) blunted transverse aortic constriction and exercise-induced cardiac myocyte hypertrophy and impaired cardiac function, demonstrating a role for ATF6 in compensatory myocyte growth. Transcript profiling and chromatin immunoprecipitation identified RHEB (Ras homologue enriched in brain) as an ATF6 target gene in the heart. RHEB is an activator of mTORC1 (mammalian/mechanistic target of rapamycin complex 1), a major inducer of protein synthesis and subsequent cell growth. Both transverse aortic constriction and exercise upregulated RHEB, activated mTORC1, and induced cardiac hypertrophy in wild type mouse hearts but not in ATF6 cKO hearts. Mechanistically, knockdown of ATF6 in neonatal rat ventricular myocytes blocked phenylephrine- and IGF1 (insulin-like growth factor 1)-mediated RHEB induction, mTORC1 activation, and myocyte growth, all of which were restored by ectopic RHEB expression. Moreover, adeno-associated virus 9- RHEB restored cardiac growth to ATF6 cKO mice subjected to transverse aortic constriction. Finally, ATF6 induced RHEB in response to growth factors, but not in response to other activators of ATF6 that do not induce growth, indicating that ATF6 target gene induction is stress specific. Compensatory cardiac hypertrophy activates ER stress and ATF6, which induces RHEB and activates mTORC1. Thus, ATF6 is a previously unrecognized link between growth stimuli and mTORC1-mediated cardiac growth.

Gallic Acid Suppresses Cardiac Hypertrophic Remodeling and Heart Failure.

Gallic acid (GA) is a dietary phenolic acid found in tea, red wine, and some plants. It exhibits anti-oxidative and anti-inflammatory activities. Recent studies have revealed that GA has beneficial effects against several cardiovascular diseases; however, whether GA attenuates pressure-overload-induced cardiac hypertrophy and the underlying mechanism remains unclear. Primary cardiomyocyte hypertrophy is stimulated with angiotensin II (Ang II). Cardiac hypertrophic remodeling is induced in mice by transverse aortic constriction (TAC). Myocardial function is evaluated by echocardiographic and hemodynamic analyses, while cardiac tissues are analyzed by histological staining. It is observed that GA significantly decreases Ang II-induced increases in cardiomyocyte size in vitro. Administration of GA in mice markedly improves TAC-induced cardiac dysfunction and attenuates pathological changes, including cardiac myocyte hypertrophy, fibrosis, inflammation, and oxidative stress. Mechanistically, GA inhibits ULK1 and activates autophagy, which induces the degradation of EGFR, gp130, and calcineurin A, thereby inhibiting the downstream signaling cascades (AKT, ERK1/2, JAK2/STAT3, and NFATc1). The results demonstrate for the first time that GA prevents myocardial hypertrophy and dysfunction via an autophagy-dependent mechanism. Thus, GA represents a promising therapeutic candidate for treating cardiac hypertrophy and heart failure.

Rationale and design of a multicentre, randomized, placebo-controlled trial of mirabegron, a Beta3-adrenergic receptor agonist on left ventricular mass and diastolic function in patients with structural heart disease Beta3-left ventricular hypertrophy (Beta3-LVH).

AimsProgressive left ventricular (LV) remodelling with cardiac myocyte hypertrophy, myocardial fibrosis, and endothelial dysfunction plays a key role in the onset and progression of heart failure with preserved ejection fraction. The Beta3‐LVH trial will test the hypothesis that the β3 adrenergic receptor agonist mirabegron will improve LV hypertrophy and diastolic function in patients with hypertensive structural heart disease at high risk for developing heart failure with preserved ejection fraction.Methods and resultsBeta3‐LVH is a randomized, placebo‐controlled, double‐blind, two‐armed, multicentre, European, parallel group study. A total of 296 patients will be randomly assigned to receive either mirabegron 50 mg daily or placebo over 12 months. The main inclusion criterion is the presence of LV hypertrophy, that is, increased LV mass index (LVMi) or increased wall thickening by echocardiography. The co‐primary endpoints are a change in LVMi by cardiac magnetic resonance imaging and a change in LV diastolic function (assessed by the E/e′ ratio). Secondary endpoints include mirabegron's effects on cardiac fibrosis, left atrial volume index, maximal exercise capacity, and laboratory markers. Two substudies will evaluate mirabegron's effect on endothelial function by pulse amplitude tonometry and brown fat activity by positron emission tomography using 17F‐fluorodeoxyglucose. Morbidity and mortality as well as safety aspects will also be assessed.ConclusionsBeta3‐LVH is the first large‐scale clinical trial to evaluate the effects of mirabegron on LVMi and diastolic function in patients with LVH. Beta3‐LVH will provide important information about the clinical course of this condition and may have significant impact on treatment strategies and future trials in these patients.

Phosphodiesterase 2A as a therapeutic target to restore cardiac neurotransmission during sympathetic hyperactivity.

Elevated levels of brain natriuretic peptide (BNP) are regarded as an early compensatory response to cardiac myocyte hypertrophy, although exogenously administered BNP shows poor clinical efficacy in heart failure and hypertension. We tested whether phosphodiesterase 2A (PDE2A), which regulates the action of BNP-activated cyclic guanosine monophosphate (cGMP), was directly involved in modulating Ca2+ handling from stellate ganglia (SG) neurons and cardiac norepinephrine (NE) release in rats and humans with an enhanced sympathetic phenotype. SG were also isolated from patients with sympathetic hyperactivity and healthy donor patients. PDE2A activity of the SG was greater in both spontaneously hypertensive rats (SHRs) and patients compared with their respective controls, whereas PDE2A mRNA was only high in SHR SG. BNP significantly reduced the magnitude of the calcium transients and ICaN in normal Wistar Kyoto (WKY) SG neurons, but not in the SHRs. cGMP levels stimulated by BNP were also attenuated in SHR SG neurons. Overexpression of PDE2A in WKY neurons recapitulated the calcium phenotype seen in SHR neurons. Functionally, BNP significantly reduced [3H]-NE release in the WKY rats, but not in the SHRs. Blockade of overexpressed PDE2A with Bay 60-7550 or overexpression of catalytically inactive PDE2A reestablished the modulatory action of BNP in SHR SG neurons. This suggests that PDE2A may be a key target in modulating the action of BNP to reduce sympathetic hyperactivity.

Load-Dependent Changes in Left Ventricular Structure and Function in a Pathophysiologically Relevant Murine Model of Reversible Heart Failure.

To better understand reverse left ventricular (LV) remodeling, we developed a murine model wherein mice develop LV remodeling after transverse aortic constriction (TAC) and a small apical myocardial infarct (MI) and undergo reverse LV remodeling after removal of the aortic band. Mice studied were subjected to sham (n=6) surgery or TAC+MI (n=12). Two weeks post-TAC+MI, 1 group underwent debanding (referred to as heart failure debanding [HF-DB] mice; n=6), whereas the aortic band remained in a second group (heart failure [HF] group; n=6). LV remodeling was evaluated by 2D echocardiography at 1 day, 2 weeks and 6 weeks post-TAC+MI. The hearts were analyzed by transcriptional profiling at 4 and 6 weeks and histologically at 6 weeks. Debanding normalized LV volumes, LV mass, and cardiac myocyte hypertrophy at 6 weeks in HF-DB mice, with no difference in myofibrillar collagen in the HF and HF-DB mice. LV ejection fraction and radial strain improved after debanding; however, both remained decreased in the HF-DB mice relative to sham and were not different from HF mice at 6 weeks. Hemodynamic unloading in the HF-DB mice was accompanied by a 35% normalization of the HF genes at 2 weeks and 80% of the HF genes at 4 weeks. Hemodynamic unloading of a pathophysiologically relevant mouse model of HF results in normalization of LV structure, incomplete recovery of LV function, and incomplete reversal of the HF transcriptional program. The HF-DB mouse model may provide novel insights into mechanisms of reverse LV remodeling.

Phosphodiesterase‐5 inhibitors “Sildenafil and Ordonafil” suppress/reverse L‐NAME induced cardiac hypertrophy in isolated rabbit heart and in phenylephrine induced hypertrophy in isolated neonatal rat myocytes

Phosphodiesterase 5 inhibitors (PDE5Is) have been shown to suppress and reverse myocardial hypertrophy induced by pressure overload. Calcineurin, a protein phosphatase, has been suggested to function as an important hypertrophic factor in cardiac myocytes and its signaling regulates the cardiac hypertrophic response in coordination with mitogen‐activated protein kinases (MAPKs). The present study investigated the effects of two PDE5Is i.e. Sildenafil (SILD) ‘ Viagra’ and Ordonafil (ORD) on N‐nitro‐L‐arginine methyl ester (L‐NAME) induced cardiac hypertrophy and assessed their ability to attenuate the activity of calcineurin, p38 and protein synthesis to exert their effect on hypertrophied cardiac myocytes.MethodsL‐NAME (10 mg/kg/day) was used to induce cardiac hypertrophy in rabbits' hearts and left ventricular thicknesses (LVTs) were measured in L‐NAME and PDE5Is treated hearts. Then, in cultured neonatal rat ventricular myocytes, Western Blot technique was used to assess the ability of these PDE5Is (different doses) to attenuate the activation of calcineurin, p38 and protein synthesis in phenylphrine‐induced ventricular hypertrophy as indicated by the cell surface area of cultured neonatal rat cardiomyocytes.ResultsL‐NAME significantly increased LVT to 6.1± 0.18 mm vs. 4.6±0.13 mm in control rabbit hearts, (p<0.05). SILD and ORD significantly regressed LVT to 4.8 ± 0.2 mm and 5.1 ± 0.1 mm, respectively (p<0.05). Phenylphrine significantly increased neonatal rat ventricular myocyte cell surface area to 131% of control values (p<0.05). This hypertrophic effect of phenylphrine was associated with significant increment in protein synthesis to 143+5% of control values, (p<0.05). Both drugs (in a dose dependent, 0.001–1 mg/L) decreased cell surface area to 95% and 90% of control value respectively, and both decreased protein synthesis to 88% of control values. Phosphatase activity of calcineurin significantly decreased from 1.93±0.09 IU to 1.15±0.05 IU after one hour and to 1.2±0.06 IU after 24 hours incubation with ORD (0.1 mg/l) incubation, (p<0.05). Whereas this activity significantly decreased after 24 hours (1.22±0.04 IU, (p<0.05)), but not after one hour (1.84±0.05 IU) of SILD (0.1 mg/l) incubation. Protein synthesis and p38 activation were significantly attenuated by both drugs (0.1 mg/L).ConclusionsThese results suggest that both SILD and ORD have the ability to reverse L‐NAME induced cardiac hypertrophy in rabbits and suppress phenylphrine induce hypertrophic myocytes. The results indicate these effects are mediated through inhibiting protein synthesis and the attenuation of calcineurin and p38 activity signaling pathways in neonatal rat ventricular myocytes which is reported in this study for the first time.Support or Funding InformationDeanship of Scientific research ‐ Jordan University of Science & TechnologyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

A biphasic model for full cycle simulation of the human heart aimed at rheumatic heart disease

Rheumatic heart disease (RHD) is identified as a serious health concern in developing countries, specifically amongst young individuals, accounting for between 250 000 and 1.4 million deaths annually. As such, attention is initially placed on the importance of the development of a cardiac analysis toolbox with functionality for pathophysiological analysis of the disease. Subsequently, in order to develop a toolbox to further the understanding of the mechanisms of the disease as linked to changes in the cytoskeletal architecture and hypertrophy of cardiac myocytes, a continuum bi-phasic model applicable to cardiac tissue is formulated based on the theory of porous media (TPM). This makes it possible to account for interactions and contributions of multiple phases of constituent materials as well as concentrations of solved components, which in computational cardiac modelling are the solid phase – the cardiac tissue – and the liquid phase – blood and interstitial fluid. Therefore, subsequent attention is paid to the cardiac model development in order to implement a sound base on which to add strain- and nutrient-driven phase transition, in addition to a nutrient phase contained within the liquid phase. To this end, based on thermodynamical restrictions, constitutive relations are proposed for stress, permeability, seepage velocity and interaction forces. The approach is implemented in the in-house computational cardiac mechanics toolbox SESKA which supports finite element as well as Element-free Galerkin-based approximations. This paper considers the passive and active non-linear elastic material behaviour of the myocardium of the left ventricle coupled with porous media theory, along with an additional coupling to the haemodynamics of the circulatory system, facilitating modelling of the full cardiac cycle. In order to illustrate the potential and efficacy of the approach with qualitative results, a human heart affected by RHD is investigated, making use of cardiovascular magnetic resonance scans taken over a period of two years to generate realistic 3D computer models.

Circulating Plasma MicroRNA-208a as Potential Biomarker of Chronic Indeterminate Phase of Chagas Disease.

Chagas cardiomyopathy is the most severe clinical manifestation of chronic Chagas disease. The disease affects most of the Latin American countries, being considered one of the leading causes of morbidity and death in the continent. The pathogenesis of Chagas cardiomyopathy is very complex, with mechanisms involving parasite-dependent cytopathy, immune-mediated myocardial damage and neurogenic disturbances. These pathological changes eventually result in cardiac myocyte hypertrophy, arrhythmias, congestive heart failure and stroke during chronic infection phase. Herein, we show that miR-208a, a microRNA that is a key factor in promoting cardiovascular dysfunction during cardiac hypertrophy processes of heart failure, has its circulating levels increased during chronic indeterminate phase when compared to cardiac (CARD) clinical forms in patients with Chagas disease. In contrast, we have not found altered serum levels of miR-34a, a microRNA known to promote pro-apoptotic role in myocardial infarction during degenerative process of cardiac injuries thus indicating intrinsic differences in the nature of the mechanisms underlying the heart failure triggered by Trypanosoma cruzi infection. Our findings support that the chronic indeterminate phase is a progressive phase involved in the genesis of chagasic cardiopathy and point out the use of plasma levels of miR-208a as candidate biomarker in risk-prediction score for the clinical prognosis of Chagas disease.

Overexpression of collagen type III in injured myocardium prevents cardiac systolic dysfunction by changing the balance of collagen distribution

ObjectiveLeft ventricular (LV) remodeling alters the contractile and relaxation properties and induces myocardial stiffness. As LV remodeling progresses, the amount of collagen type III (Col3) is gradually decreased, being replaced by collagen type I (Col1). We evaluated whether Col3 overexpression improved cardiac function and remodeling in a rat with ischemic cardiomyopathy (ICM). We also investigated the functional motif and mechanism of thrombin-cleaved N-terminal osteopontin (N-OPN) on cardiac remodeling. MethodsThe rats with ICM were divided into 3 groups: ligation only (Control) group and groups transplanted with nontransfected fibroblast sheets (normal Fb group) or with Col3-secretory fibroblast sheets (Col3 Fb group). A gelatin hydrogel containing the N-terminal fragment (N-OPN), N-OPN lacking the SVVYGLR sequence (⊿SV), the Arg-Gly-Asp (RGD) sequence (⊿RGD), RGD and SVVYGLR sequences (⊿RGD-SV), SVVYGLR alone (SV), or a random SV peptide was implanted into an ICM model rat. ResultsThe Col3 Fb group exhibited significantly attenuated LV systolic dysfunction. LV dilatation, myocyte hypertrophy, and LV fibrosis at the infarcted area were also attenuated by Col3 Fb implantation. Furthermore, N-OPN, ⊿RGD, and SV peptide suppressed the depression of cardiac function, LV dilatation, and myocyte hypertrophy, and also induced increased Col3 expression and reduction in the ratio of Col1 to Col3 in the infarcted and border areas. ConclusionsOverexpression of Col3 improved cardiac function by changing the balance of collagen distribution in LV remodeling. The SVVYGLR motif of the thrombin-cleaved N-OPN and SV peptide attenuated cardiac dysfunction by increasing Col3 and changing the pattern of collagen balance in the impaired area.

Hamartoma of mature cardiac myocytes: Report of a rare case with review of literature

Hamartoma of mature cardiac myocyte is a benign tumor of the heart. They are characterized by haphazardly arranged hypertrophic cardiac myocytes with variable amounts of intervening fibrosis, mature adipose tissue along with irregularly distributed variable-sized vascular channels. The case presented here had a hyperechoic mass in the left ventricle arising from the midinferior and posterior interventricular septum, infiltrating into the adjacent myocardium. The MRI , histopathology findings and the clinical outcome is presented.

Ursolic acid ameliorates oxidative stress, inflammation and fibrosis in diabetic cardiomyopathy rats

Diabetic cardiomyopathy is a major and severe cardiovascular complication of diabetes mellitus. Ursolic acid, a pentacyclic triterpene compound widespread in fruits and plants, performs a variety of pharmacological activities including lowering blood glucose, anti-oxidation, anti-inflammation and anti-fibrosis. Our present study aimed to investigate the cardioprotective effects of ursolic acid on diabetic cardiomyopathy rats and uncover its underlying mechanism. Diabetes mellitus was induced by a single injection of STZ-only (40 mg/ kg, i.v.) in male SD rats. Animals were divided into three groups (n=10): control group (normal saline, i.g.), diabetic group (normal saline, i.g.) and diabetic+ursolic acid group (35 mg/kg UA + normal saline, i.g.). Rats were administered for 8 weeks from 5th to 12th week. After the last administration, cardiac function was evaluated; HWI was calculated; FBG, CK, LDH in serum and SOD, MDA in cardiac tissue were detected. HE staining and Masson trichrome staining were employed to observe pathological alterations. Immunohistochemistry and western blotting were taken to determine the expression levels of TNF-α, MCP-1, TGF-β1 and MMP-2 in the heart. The results dramatically showed increased levels of FBG, CK, LDH, MDA and a decreased activity of SOD in diabetic group, in which left ventricular dysfunction, cardiac myocytes hypertrophy, inflammatory cell infiltration and myocardial interstitial fibrosis had also been found. What's more, the expressions of TNF-α, MCP-1 and TGF-β1 were significantly up-regulated and the expression of MMP-2 was markedly down-regulated in myocardium. Interestingly, treatment with ursolic acid remarkably ameliorated these changes. Collectively, our study strongly showed that ursolic acid is capable of improving the cardiac structure and function in STZ-induced diabetic cardiomyopathy rats by attenuating oxidative stress, inflammation and fibrosis.

Cardiac myocyte miR-29 promotes pathological remodeling of the heart by activating Wnt signaling

Chronic cardiac stress induces pathologic hypertrophy and fibrosis of the myocardium. The microRNA-29 (miR-29) family has been found to prevent excess collagen expression in various organs, particularly through its function in fibroblasts. Here, we show that miR-29 promotes pathologic hypertrophy of cardiac myocytes and overall cardiac dysfunction. In a mouse model of cardiac pressure overload, global genetic deletion of miR-29 or antimiR-29 infusion prevents cardiac hypertrophy and fibrosis and improves cardiac function. Targeted deletion of miR-29 in cardiac myocytes in vivo also prevents cardiac hypertrophy and fibrosis, indicating that the function of miR-29 in cardiac myocytes dominates over that in non-myocyte cell types. Mechanistically, we found cardiac myocyte miR-29 to de-repress Wnt signaling by directly targeting four pathway factors. Our data suggests that, cell- or tissue-specific antimiR-29 delivery may have therapeutic value for pathological cardiac remodeling and fibrosis.

Estrogen receptor beta maintains expression of KLF15 to prevent cardiac myocyte hypertrophy in female rodents.

Maintaining a healthy, anti-hypertrophic state in the heart prevents progression to cardiac failure. In humans, angiotensin II (AngII) indirectly and directly stimulates hypertrophy and progression, while estrogens acting through estrogen receptor beta (ERβ) inhibit these AngII actions. The KLF15 transcription factor has been purported to provide anti-hypertrophic action. In cultured neonatal rat cardiomyocytes, we found AngII inhibited KLF1 expression and nuclear localization, substantially prevented by estradiol (E2) or β-LGND2 (β-LGND2), an ERβ agonist. AngII stimulation of transforming growth factor beta expression in the myocytes activated p38α kinase via TAK1 kinase, inhibiting KLF15 expression. All was comparably reduced by E2 or β-LGND2. Knockdown of KLF15 in the myocytes induced myocyte hypertrophy and limited the anti-hypertrophic actions of E2 and β-LGND2. Key aspects were confirmed in an in-vivo model of cardiac hypertrophy. Our findings define additional anti-hypertrophic effects of ERβ supporting testing specific receptor agonists in humans to prevent progression of cardiac disease.

Apoptosis signal-regulating kinase 1 inhibition attenuates cardiac hypertrophy and cardiorenal fibrosis induced by uremic toxins: Implications for cardiorenal syndrome

Intracellular accumulation of protein-bound uremic toxins in the setting of cardiorenal syndrome leads to adverse effects on cardiorenal cellular functions, where cardiac hypertrophy and cardiorenal fibrosis are the hallmarks. In this study, we sought to determine if Apoptosis Signal-Regulated Kinase 1 (ASK1), an upstream regulator of cellular stress response, mediates cardiac hypertrophy and cardiorenal fibrosis induced by indoxyl sulfate (IS) and p-cresol sulfate (PCS) in vitro, and whether ASK1 inhibition is beneficial to ameliorate these cellular effects. PCS augmented cardiac myocyte hypertrophy and fibroblast collagen synthesis (as determined by 3H-leucine and 3H-proline incorporation, respectively), similar to our previous finding with IS. IS and PCS also increased collagen synthesis of proximal tubular cells and renal mesangial cells. Pro-hypertrophic (α-skeletal muscle actin and β-MHC) and pro-fibrotic genes (TGF-β1 and ctgf) were induced by both IS and PCS. Western blot analyses revealed the activation of ASK1 and downstream mitogen activated protein kinases (MAPKs) (p38MAPK and ERK1/2) as well as nuclear factor-kappa B (NF-κB) by IS and PCS. ASK1, OAT1/3, ERK1/2 and p38MAPK inhibitors suppressed all these effects. In summary, IS and PCS exhibit pro-hypertrophic and pro-fibrotic properties, at least in part, via the activation of ASK1 and its downstream pathways. ASK1 inhibitor is an effective therapeutic agent to alleviate protein-bound uremic toxin-induced cardiac hypertrophy and cardiorenal fibrosis in vitro, and may be translated further for cardiorenal syndrome therapy.

Ca2+/Calmodulin-Dependent Protein Kinase II and Androgen Signaling Pathways Modulate MEF2 Activity in Testosterone-Induced Cardiac Myocyte Hypertrophy.

Testosterone is known to induce cardiac hypertrophy through androgen receptor (AR)-dependent and -independent pathways, but the molecular underpinnings of the androgen action remain poorly understood. Previous work has shown that Ca2+/calmodulin-dependent protein kinase II (CaMKII) and myocyte-enhancer factor 2 (MEF2) play key roles in promoting cardiac myocyte growth. In order to gain mechanistic insights into the action of androgens on the heart, we investigated how testosterone affects CaMKII and MEF2 in cardiac myocyte hypertrophy by performing studies on cultured rat cardiac myocytes and hearts obtained from adult male orchiectomized (ORX) rats. In cardiac myocytes, MEF2 activity was monitored using a luciferase reporter plasmid, and the effects of CaMKII and AR signaling pathways on MEF2C were examined by using siRNAs and pharmacological inhibitors targeting these two pathways. In the in vivo studies, ORX rats were randomly assigned to groups that were administered vehicle or testosterone (125 mg⋅kg-1⋅week-1) for 5 weeks, and plasma testosterone concentrations were determined using ELISA. Cardiac hypertrophy was evaluated by measuring well-characterized hypertrophy markers. Moreover, western blotting was used to assess CaMKII and phospholamban (PLN) phosphorylation, and MEF2C and AR protein levels in extracts of left-ventricle tissue from control and testosterone-treated ORX rats. Whereas testosterone treatment increased the phosphorylation levels of CaMKII (Thr286) and phospholambam (PLN) (Thr17) in cardiac myocytes in a time- and concentration-dependent manner, testosterone-induced MEF2 activity and cardiac myocyte hypertrophy were prevented upon inhibition of CaMKII, MEF2C, and AR signaling pathways. Notably, in the hypertrophied hearts obtained from testosterone-administered ORX rats, both CaMKII and PLN phosphorylation levels and AR and MEF2 protein levels were increased. Thus, this study presents the first evidence indicating that testosterone activates MEF2 through CaMKII and AR signaling. Our findings suggest that an orchestrated mechanism of action involving signal transduction and transcription pathways underlies testosterone-induced cardiac myocyte hypertrophy.

G protein-coupled receptor kinase 2 promotes cardiac hypertrophy.

The increase in protein activity and upregulation of G-protein coupled receptor kinase 2 (GRK2) is a hallmark of cardiac stress and heart failure. Inhibition of GRK2 improved cardiac function and survival and diminished cardiac remodeling in various animal heart failure models. The aim of the present study was to investigate the effects of GRK2 on cardiac hypertrophy and dissect potential molecular mechanisms. In mice we observed increased GRK2 mRNA and protein levels following transverse aortic constriction (TAC). Conditional GRK2 knockout mice showed attenuated hypertrophic response with preserved ventricular geometry 6 weeks after TAC operation compared to wild-type animals. In isolated neonatal rat ventricular cardiac myocytes stimulation with angiotensin II and phenylephrine enhanced GRK2 expression leading to enhanced signaling via protein kinase B (PKB or Akt), consecutively inhibiting glycogen synthase kinase 3 beta (GSK3β), such promoting nuclear accumulation and activation of nuclear factor of activated T-cells (NFAT). Cardiac myocyte hypertrophy induced by in vitro GRK2 overexpression increased the cytosolic interaction of GRK2 and phosphoinositide 3-kinase γ (PI3Kγ). Moreover, inhibition of PI3Kγ as well as GRK2 knock down prevented Akt activation resulting in halted NFAT activity and reduced cardiac myocyte hypertrophy. Our data show that enhanced GRK2 expression triggers cardiac hypertrophy by GRK2-PI3Kγ mediated Akt phosphorylation and subsequent inactivation of GSK3β, resulting in enhanced NFAT activity.